Rapid Homemade Reverse Transcriptase PCR Detection and Phylogenetic Analysis for SARS-CoV-۲ Based on E and M Genes

Background & Objective: Coronavirus disease ۲۰۱۹ spreads worldwide and needs detection systems capable of rapid diagnostic of this virus (SARS-CoV-۲). The aim of this study is to design the homemade RT-PCR method for the Detection and phylogenetic analysis of this virus Material & Methods: The genes selected for diagnosis were E and M genes for this virus. PCR product was cloned in pTZ۵۷R/T plasmid for preparation of positive control. In order to determine the sensitivity of this molecular method, the genes mentioned in the clone pTZ۵۷R/T vector and the Limit of detection (LOD) the genes were determined and phylogenetic analysis was performed using partial E and M gene sequences. Results: PCR product was observed for E and M genes ۱۵۶ and ۵۴۷ bp on the Agarose gel. The LOD of the E and M gene was ۶۰ and ۸۲ copies. There was also a positive response to the samples of patients who were positive by other methods. Conclusions: Since this virus is considered to be the cause of a pandemic in different countries all over the world, the present study is very important as a method of rapid and low-cost molecular diagnosis for monitoring this virus. Phylogenetic analysis is necessary for epidemiological studies for the control and prevention of the disease.

کلیدواژه ها:

RT-PCR ، Covid-۱۹ ، Detection ، LOD ، Phylogenetic Analysis